Tarsi of Heliothine Moths Contain Aldehydes and Butyrate

TARSI OF HELIOTHINE MOTHS CONTAIN ALDEHYDES AND BUTYRATE

ESTERS AS POTENTIAL PHEROMONES

Journal of Chemical Ecology

MAN-YEON CHOI1, SEUNG-JOON AHN2, KYE-CHUNG PARK3, ROBERT VANDER

MEER4, RING T. CARDÉ5, RUSSELL JURENKA6,*

1USDA-ARS, Horticultural Crops Research Laboratory, 3420 NW Orchard Avenue, Corvallis, OR 97330, USA; 2Department of Entomology, Max Planck Institute for Chemical Ecology, 07745 Jena, Germany; 3Horticulture and Food Research Institute Gerald Street, 8152 Lincoln, New Zealand; 4USDA-ARS, Center for medical, agricultural and veterinary entomology, Gainesville, FL 32608, USA; 5 Department of Entomology, University of California, Riverside, CA 92521; 6434 Science II, Department of Entomology, Iowa State University, Ames, IA 50011, USA

*Correspondence to Russell Jurenka ()

Supplemental Material

Table S1.Sex pheromone compoundss found in female pheromone glands or legs of both malesand females of Helicoverpa and Heliothis moths.

Pheromone gland
major component / fore-leg / mid-leg / hind-leg
Helicoverpaarmigera / Z11-16:Ald / 16:Ald / 16:Ald / 16:Ald
Helicoverpaassulta / Z9-16:Ald / 16:Ald / 16:Ald / 16:Ald
Helicoverpazea / Z11-16:Ald / 16:Ald / 16:Ald / 16:Ald
Heliothisvirescens / Z11-16:Ald / 16:Ald / 16:Ald / 16:Ald

Table S2. The identified major and the most important minor sex pheromone components of female heliothine moths. The 16:Ald has been identified as a compound present in pheromone glands of these heliothine species.

/ major / minor / present
Helicoverpaarmigera / Z11-16:Ald / Z9-16:Ald / 16:Ald
Helicoverpaassulta / Z9-16:Ald / Z11-16:Ald / 16:Ald
Helicoverpagelotopoeon / Z9-16:Ald / 16:Ald / 16:Ald
Helicoverpazea / Z11-16:Ald / Z9-16:Ald / 16:Ald
Heliothis belladonna / Z11-16:Ald / Z9-16:Ald / 16:Ald
Heliothis maritime / Z11-16:Ald / Z9-16:Ald / 16:Ald
Heliothispeltigera / Z11-16:Ald / Z9-16:Ald / 16:Ald
Heliothisphloxiphaga / Z11-16:Ald / Z9-16:Ald / 16:Ald
Heliothissubflexa / Z11-16:Ald / Z9-16:Ald / 16:Ald
Heliothisvirescens / Z11-16:Ald / Z9-16:Ald / 16:Ald
Abundance

16:Ald

5782

43

96

68

196222

20406080 100 120 140 160 180 200 220 240 m/z

IS / 18:Ald
27:HC
14 / 16 / 18 / 20 / 22 / 24 / 26 / 28
Retention time (min)

Figure S1 Partial GC/MS total ion current chromatograms of hexane extracts from the tarsi of Helicoverpaarmigera males. Relevant peaks are labeled. The mass spectra of 16:Ald is shown in the inset. IS = internal standard which represents 500 ng of Z9-14:Ald. 27:HC = heptacosane.

400 / ab / scotophase
ng/moth / 300 / b / photophase
a
16:Ald, / a / a
200 / a / a / a
a
a
100
0 / 1 / 2 / 3 / 4
Number of days after emergence

Figure S2 Amount of 16:Ald found on legs without tarsi of maleHelicoverpazeaduring the scotophaseor photophase at the indicated times after adult emergence. Bars indicate mean + SEM; bars with the same letters are not statistically different at P≤ 0.05 using one-way ANOVA followed byTukey’s multiple comparison test; N ≥ 5

ng/moth16:Ald,
400 / scotophase
300 / photophase
200 / a
100 / b / b / b / b / b
b / b
b / b
0 / 1 / 2 / 3 / 4
Number of days after emergence

Figure S3 Amount of 16:Ald found on legs without tarsi taken from femaleHelicoverpazeaduring thescotophase or photophase at the indicated times after emergence. Bars indicate mean + SEM; bars with the same letters are not statistically different at P ≤ 0.05 using one-way ANOVAfollowed by Tukey’s multiple comparison test; N ≥ 5

400 / male
ng/moth / 300 / b / female
a / b
16:Ald, / 200 / a
ab
100 / a / a / a
24 / 48 / 72 / 96
Time after decapitation, h

Figure S4 Amount of 16:Ald found on legs without tarsi taken fromHelicoverpazeadecapitated males orfemales at the indicated times post-decapitation. Bars indicate mean + SEM; bars with the same letters are not statistically different at P ≤ 0.05 using one-way ANOVA followed by Tukey’smultiple comparison test; N ≥ 5

16:Ald

18:Ald

Relative abundance
15 / 16 / 17 / 18 / 19

Retention Time, min

Figure S5Partial GC/MS total ion current chromatograms from the solid phase microextraction of the headspace collected from tarsi of Helicoverpazea males. Relevant peaks are labeled.

A / Male tarsi
hydrocarbons / 31-1
butyrate esters
BuO-27
response / BuO-29 (1-4)
3
29-2 / 2
detector / 29-1
1
BuO-25 / BuO-29
Relative / 27 / 31-2
4
29 / 33-2
25 / BuO-31 (1)
23 / 24 / 25 / 26 / 27 / 28 / 29 / 30 / 31 / 32

Retention Time, min

Figure S6 A. Partial GC/MS total ion current chromatograms of purified hydrocarbons and butyrate esters(BuO) extracted from the tarsi of Helicoverpazea. The peaks of the hydrocarbon profile are labeled with the chain length followed by the number of methyl branches. The peaks of the butyrate esters are labeled with the chain number. A series of butyrate esters with retention times after the BuO-29 peaks (labeled 1 -4) are indicated. These butyrate esters could have methyl branches. The mass spectra suggest an increase in the molecular weight of peaks labeled BuO-29 (1-4) as shown in Fig. S6B.

71BuO-25

57

89 / 111 / 350
125 / 199 / 267
60 / 80 / 100 / 120 / 140 / 160 / 180 / 200 / 220 / 240 / 260 / 280 / 300 / 320 / 340 / 360

71

BuO-29

89 / 111
139 / 348 / 406
60 / 80 / 100 / 120 / 140 / 160 / 180 / 200 / 220 / 240 / 260 / 280 / 300 / 320 / 340 / 360 / 380 / 400

71BuO-29 (2)

55 / 111
89
153
420

406080100120140160180200220240260280300320340360380400420

71BuO-27

89 / 111 / 378
139 / 213
40 / 60 / 80 / 100 / 120 / 140 / 160 / 180 / 200 / 220 / 240 / 260 / 280 / 300 / 320 / 340 / 360 / 380
71 / BuO-29 (1)
57
97 / 139 155 / 181 / 348 / 365 / 420
40 / 60 / 80 / 100 / 120 / 140 / 160 / 180 / 200 / 220 / 240 / 260 / 280 / 300 / 320 / 340 / 360 / 380 / 400 / 420

71

BuO-29 (3)

57
89
111
139 / 420

406080100120140160180200220240260280300320340360380400420

71 / BuO-29 (4)

7 1

BuO-31 (1)

57
89 / 111 / 139 / 199 / 251 / 340 / 434
40 / 60 / 80 / 100 / 120 / 140 / 160 / 180 / 200 / 220 / 240 / 260 / 280 / 300 / 320 / 340 / 360 / 380 / 400 / 420 / 440
57
89 / 111 / 195 / 278 / 448
40 / 60 / 80 / 100 / 120 / 140 / 160 / 180 / 200 / 220 / 240 / 260 / 280 / 300 / 320 / 340 / 360 / 380 / 400 / 420 / 440

Figure S6B Mass spectra of the BuO- 29, BuO- 29 (1-4), and BuO- 31 (1) labeled peaks in Fig. S6A. The increase of 14 mass units in BuO - 29 (1- 3) and 28 mass units in BuO- 29 (4) compared to BuO-29 is indicative of the presence of one methyl or two methyl groups, respectively. The BuO-31 (1) mass spectra indicates a butyl ester with 31 carbons in the chain and a methyl group.

A
*
*
response / **
detector / B
Relative

*

* *

23242526272829303132

Retention Time, min

Figure S7 Representative partial GC/MS total ion current chromatograms of hydrocarbons and butyrate esters extracted from the mesothoracic tarsi (A) and the femur and tibia of the mesothoracic leg (B) of Helicoverpazea. The butyrate esters are labeled with an asterisk.

response / *
detector / * / **
Relative / *
*
*
23 / 24 / 25 / 26 / 27 / 28 / 29 / 30 / 31 / 32

Retention Time, min

Figure S8 Representative partial GC/MS total ion current chromatogram of the hydrocarbons and butyrate esters extracted from a Petri dish after 4 Helicoverpazea males were kept in the dish for 12 h. The asterisk above the peak indicates the butyrate esters.